Anti-NY-ESO-1 scFv h(CD28-CD3ζ) CART, pCDCAR1 vector

All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

Details

Target
NY-ESO-1

Targeting Cell Type
T cell

Targeting Diseases
Metastatic melanoma; synovial cell sarcoma

Generation
Second

Vector Name
pCDCAR1

Vector Length
8kb

Vector Type
Lentiviral

Receptor Construction
scFv-CD28-CD3ζ

Discription of Signaling Cassetes
CD28
CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co-stimulatorysignals required for T cell activation and survival. CD28 is the receptor for CD80 (B7.1) and CD86 (B7.2) proteins which are expressed on antigen-presenting cells(APC). CD28 modulates the primary TCR/CD3ζ signal in a different fashion than the late costimulatory elements OX40 and 4-1BB. CD28 enhances the expression of downstream regulators that impact on T-cell proliferation, death, differentiation, and effector functions. CAR+ T cells containing the CD28 endodomain showed strikingly enhanced sustained T cell activation, growth, survival. And CD28 results in a brightly expressed, stable receptor as the transmembrane domain. Including CD28 costimulatory domains in CARs led to enhanced anti-malignancy efficacy.
CD3ζ
CD3ζ, also known as T-cell receptor zeta, which together with T-cell receptor and CD3γ, δ , ε chain, forms the TCR-CD3 complex. ζ was expressed independently from the complex. The zeta chain plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. CD3-zeta,which contains 3 ITAMs, is the most commonly used endodomain component of CARs. It transmits an activation signal to the T cell after antigen is bound. CD3-zeta may not provide a fully competent activation signal and additional co-stimulatory signaling is needed. For example, chimeric CD28 and OX40 can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.

Target

Customize Your CAR Products

Published Data

Complete CAR data ELISA

Fig.1 Antibody titration of positive clone mAb 2D2 via ELISA.

CAR Construction : 2D2 scFv-BBζ

Fig.1 Antibody titration of positive clone mAb 2D2 via ELISA.

Selection of scFvs specific for HLA-A2/NY-ESO-1 complex.

Liu, X., Xu, Y., Xiong, W., Yin, B., Huang, Y., Chu, J., ... & Wang, R. (2022). Development of a TCR-like antibody and chimeric antigen receptor against NY-ESO-1/HLA-A2 for cancer immunotherapy. Journal for immunotherapy of cancer, 10(3).

Complete CAR data BLI

Fig.2 Binding affinity of mAb 2D2 to the HLA-A2/NY-ESO-1157-165.

CAR Construction : 2D2 scFv-BBζ

Fig.2 Binding affinity of mAb 2D2 to the HLA-A2/NY-ESO-1157-165.

The complex measured by the biolayer interferometry (BLI) on an Octet instrument.

Complete CAR data FCM

Fig.3 Surface expression of CAR detected by HLA-A2/NY-ESO-1157-165 complex.

CAR Construction : 2D2 scFv-BBζ

Fig.3 Surface expression of CAR detected by HLA-A2/NY-ESO-1157-165 complex.

2D2-CAR T cells specifically recognize and lyze HLA-A2+/NY-ESO-1+cells in vitro.

Complete CAR data Cyt

Fig.4 LDH cytotoxicity assay of 2D2-CAR T cells.

CAR Construction : 2D2 scFv-BBζ

Fig.4 LDH cytotoxicity assay of 2D2-CAR T cells.

2D2-CAR T cells were cocultured with T2 cells that were pulsed with 20µg/mL NY-ESO-1 or CMV peptide for 1 hour for 4 hours.

Complete CAR data FuncS

Fig.5 2D2-CAR T cells impair tumor growth and prolong the survival of mice bearing triple-negative breast cancer in vivo.

CAR Construction : 2D2 scFv-BBζ

Fig.5 2D2-CAR T cells impair tumor growth and prolong the survival of mice bearing triple-negative breast cancer in vivo.

Representative survival analysis of MBA-MD-231-NY-ESO-1 bearing mice treated with 2D2-CAR T cells compared with the control group.

Complete CAR data ELISA

Fig.6 IFN-γcytokine release measured by ELISA.

CAR Construction : 2D2 scFv-BBζ

Fig.6 IFN-γcytokine release measured by ELISA.

2D2-CAR T cells were cocultured with target cells with a 10:1 (E: T) ratio overnight.

Complete CAR data IHC

Fig.7 Confocal imaging of mAb 2D2 staining in NY-ESO1157-165 peptide-pulsed HEK 293T.

CAR Construction : 2D2 scFv-BBζ

Fig.7 Confocal imaging of mAb 2D2 staining in NY-ESO1157-165 peptide-pulsed HEK 293T.

HEK 293T cells were pulsed with 20µg/mL NY-ESO-1157-165 or CMV pp65495-503 peptide for 1 hour.

More Published Data More Published Data

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